The study on the effect of Compound Chinese Medicine on lumbar ligament cells

• 1、Bioengineering College,Chongqing University,Chongqing 400044

Abstract：Objective: To investigate the effects of Compound Traditional Chinese Medicine (CTCM) on the expression of extracellular matrix proteins, degrading enzymes and endogenous growth factors in Interspinous LiThe study on the effect of Compound Chinese Medicine on lumbar ligament cellsgament Fibroblasts (ILFs) of rabbits. Methods: The ILFs of New Zealand white rabbits were isolated and cultured in vitro, and the third-generation ILFs were randomly divided into three groups: the normal control group, the CTCM group (200 μg / mL), and the Shujinjianwan group (S group, 30 μg / mL). Cell viability was measured after in vitro culture for different periods of time. The gene level of collagenⅠ, collagen Ⅲ, elastin, mTOR, growth factors and MMP-2, MMP-9 in different groups of cells was detected by RT-PCR. Immunofluorescence staining was used to semi-quantitatively detect the expression of collagenⅠ, collagen Ⅲ and elastin in different groups of cells and observe the morphology of collagen assembly. Besides, Western blot was used to quantitatively detect the expression of collagen, elastin and phosphorylated mTOR. Results: PCR results showed that the CTCM group and the S group could significantly up-regulate the gene level of type I, III collagen, elastin, and mTOR related to the PI3K / AKT signaling pathway after 3 days, and similar to EGF, bFGF, and MGF. Immunofluorescence staining results showed that collagen I, collagen III and elastin in the CTCM group and the S group increased significantly on the 3rd day, but no obvious changes were observed in the collagen assembly morphology. Western blot results showed that the expression of type I, III collagen, elastin, and phosphorylated mTOR of CTCM group and the S group were significantly up-regulated compared to normal group on the 3rd day and 4th day. In addition, the CTCM group and the S group down-regulated the expression of matrix metalloproteinase on the 1st day, and up-regulated on the 3rd and 5th days. Conclusion: CTCM promote the expression of ILFs extracellular matrix proteins and endogenous growth factors, and down-regulate matrix metalloproteinases in the early stage of repair, in addition, its significant up-regulation in the late stage of matrix shaping, which is conducive to the repair of ligaments.

Keywords： Compound Chinese medicine Lumbar ligament cells Collagen Growth factor Matrix metalloproteinase?????